Antibacterial, Anti-Biofilm, and Anti-Inflammatory Properties of Gelatin-Chitosan-Moringa-Biopolymer-Based Wound Dressings towards Staphylococcus aureus and Escherichia coli.

In contemporary times, the sustained aspiration of bioengineering and biomedical applications is the progressive advancement of materials characterized by biocompatibility and biodegradability. The investigation of the potential applications of polymers as natural and non-hazardous materials has placed significant emphasis on their physicochemical properties. Thus, this study was designed to investigate the potential of gelatin-chitosan-moringa leaf extract (G-CH-M) as a novel biomaterial for biomedical applications. The wound-dressing G-CH-M biopolymer was synthesized and characterized. The blood haemolysis, anti-inflammatory, antioxidant, and antibacterial activities of the biopolymer were investigated against Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli) bacterial isolates. Our results showed that S. aureus swarming motility was drastically affected. However, the biopolymer had no significant effect on the swarming motility of E. coli. In addition, the biopolymer showed high antibacterial capacities, especially against S. aureus. Plasmid DNA was observed to be effectively protected from oxidative stresses by the biopolymer. Furthermore, the biopolymer exhibited greatly suppressed haemolysis (lower than 2%), notwithstanding the elevated concentration of 50 mg/mL. These results indicated that this novel biopolymer formulation could be further developed for wound care and contamination prevention.

Inhibitory potential of natural plant extracts against Escherichia coli strain isolated from diarrheic camel calves.

Background: Camel calf's diarrhea is considered the chief economic loss in the camelid population. There is currently no vaccine licensed to prevent colibacillosis in camel calves. The new era of bacterial antibiotic resistance explains the treatment failure and the high mortality and morbidity associated with the disease. Current protective treatments have thus far limited efficacy and need to be replaced. Due to their antimicrobial properties and safety, natural products are recently finding a capital role in infection management. Aims: The current study explores Escherichia coli F17 susceptibility as a clinical strain isolated from diarrheic camel calves to a wide panel of natural products. Methods: Agar diffusion method, integrity of cell membrane, hydrophobicity of bacterial surface, biofilm assays, and motility were used to evaluate the antibacterial activity of Coffea, Retama raetam, Moringa oleifera, Juniperus phoenicea, Uritica dioica, Camellia sinensis, Lavandula angustifolia, and Cuminum cyminum extracts against isolated bacteria. Results: Interestingly, all eight tested extracts have the damaging ability of E. coli F17's cell membrane and cause the nucleic acid release after 12 hours. Escherichia coli F17 strain has the surface of hydrophobicity which changed after contact with extracts of the plant. Moreover, the motility of the studied bacteria changed after exposure to all plant extracts. Conclusion: This study demonstrated that all extracts, exempt U. dioica, can remove up to 50% biofilm of E. coli biomass as compared with the control. Natural extracts can be used as potential antimicrobial agents to mitigate diarrhea in camel calves.

Perspective on therapeutic and diagnostic potential of camel nanobodies for coronavirus disease-19 (COVID-19).

In this paper, we focus on the camelid nanobodies as a revolutionary therapy that can guide efforts to discover new drugs for Coronavirus disease (COVID-19). The small size property makes nanobodies capable of penetrating efficiently into tissues and recognizing cryptic antigens. Strong antigen affinity and stability in the gastrointestinal tract allow them to be used via oral administration. In fact, the use of nanobodies as inhalant can be directly delivered to the target organ, conferring high pulmonary drug concentrations and low systemic drug concentrations and minimal systemic side effects. For that, nanobodies are referred as a class of next-generation antibodies. Nanobodies permit the construction of multivalent formats that may achieve ultra-high neutralization potency and then may prevent mutational escape and can neutralize a wide range of SARS-CoV-2 variants. Due to their distinctive characteristics, nanobodies can be of great use in the development of promising treatment or preventive strategies against SARS-CoV-2 infection. In this review, the state-of-the-art of camel nanobodies design strategies against the virus including SARS-CoV-2 are critically summarized. The application of general nanotechnology was also discussed to mitigate and control emerging SARS-CoV-2 infection.

Characterization and antimicrobial susceptibility of Escherichia coli isolated from healthy farm animals in Tunisia.

Healthy animals can constitute a reservoir for Escherichia coli potentially dangerous for humans. Our objectives were to investigate virulence genes in E. coli isolated from healthy animals in southern Tunisia and to determine their resistance to antimicrobials of high importance in humans and animals. 126 fecal samples were collected from healthy animals (cattle, sheep, goats, chicken, camel, bustard and rabbit) and assayed by PCR for virulence genes and by disk diffusion for antimicrobial resistance. STEC were isolated most frequently from goats (27.7%), sheep (20%) and cattle (14.2%). ExPEC prevalence of iucD (41.6%), papC (27.7%), sfa (13.8%), afa8 (13.8%) and iron (72.2%) was highest in camels. Prevalence of the ExPEC associated genes iss and cnf and the EPEC defining gene eae was highest in rabbits (53.3, 13.3, and 53.3%, respectively). The genes defining enterotoxigenic, enteroinvasive and enteroaggregative E. coli were not detected and faeG was found only in camels (5.5%). The most common phylogenetic groups were B1 (54.5%) and B2 (16.6%). Virulence gene profiles varied greatly between animal species. Overall, antimicrobial resistance was not highly prevalent, the highest resistance being observed against tetracycline, 43.9%.

Construction of a Nanobodies Phage Display Library From an Escherichia coli Immunized Dromedary.

BACKGROUND: Diarrhea caused by Escherichia coli is a major cause of morbidity and mortality in young animals. Few treatment options are available, mainly antibiotic therapy increasingly limited by resistance to commonly used drugs. OBJECTIVES: The aim of this work was to develop immunotherapy based on the use of camel VHH antibody fragments, or nanobodies, to target pathogenic E. coli surface antigens. MATERIAL AND METHODS: We immunized a camel with a killed strain we had previously isolated from a diarrheic camel calf and identified as expressing the F17 fimbriae antigen. RESULTS: The immunized animal developed an anti-E.coli immune response including heavy-chain antibodies. Lymphocytes from this animal were purified and RNA isolated to create a VHH library by phage display with a size of about 109 individual transformants. Panning on live E. coli cells resulted in the isolation of VHH fragments specific to the cell surface antigens. CONCLUSION: The identification of these antigens can lead to the development of new diagnostic and therapeutic tools against diarrhea.

Screening for fecal presence of colistin-resistant Escherichia coli and mcr-1 and mcr-2 genes in camel-calves in southern Tunisia.

Camels (Camelus dromedarius) are known to harbor multidrug resistant Gram-negative bacteria and to be involved in the transmission of various microorganisms to humans. Data on the occurrence of colistin resistant Escherichia coli as well as mobilized colistin resistance (mcr) genes in camels are lacking. We investigated the presence of colistin resistance and mcr (1-2) genes in E. coli from the feces of camels in Tunisia. Presumptive E. coli isolates from camel-calves in southern Tunisia were qualitatively screened for growth on Mueller-Hinton agar supplemented with 2 mg/L of colistin. The minimal inhibitory concentration of colistin was determined for isolates growing on this medium. All isolates were screened for the presence of the mcr-1 and mcr-2 genes by polymerase chain reaction without detecting any of these genes. However, one isolate was confirmed resistant to colistin and further testing of this isolate revealed it to be Enterobacter cloacae. Our study demonstrated absence of colistin resistance and of the mcr-1 and mcr-2 genes in E. coli isolated from camel feces in southern Tunisia. Thus, there is no evidence that camels represent a major source of mcr genes contamination for the local population or for tourists visiting southern Tunisia.

Antimicrobial resistance and molecular characterization of virulence genes, phylogenetic groups of Escherichia coli isolated from diarrheic and healthy camel-calves in Tunisia.

This study was conducted to determine the prevalence of virulence genes, serogroups, antimicrobial resistance and phylogenetic groups of Escherichia coli strains isolated from diarrheic and healthy camel calves in Tunisia. From 120 fecal samples (62 healthy and 58 diarrheic camel calves aged less than 3 months), 70 E. coli isolates (53 from diarrheic herds and 17 from healthy herds) were examined by PCR for detection of the virulence genes associated with pathogenic E. coli in animals. A significantly greater frequency of the f17 gene was observed in individual camels and in herds with diarrhea, this gene being found in 44.7% and 41.5% of isolates from camels and herds with diarrhea versus 22.5% and 11.7% in camels (p=0.05) and herds without diarrhea (p=0.02). The aida, cnf1/2, f18, stx2 and paa genes were found only in isolates from camels with diarrhea, although at a low prevalence, 1.8%, 3.7%, 1.8%, 3.7% and 11.3%, respectively. Prevalence of afa8, cdtB, eae, east1, iroN, iss, kpsMTII, paa, sfa, tsh and papC genes did not differ significantly between herds with or without diarrhea. Genes coding for faeG, fanC, f41, estI, estII, CS31a and eltA were not detected in any isolates. All isolates were sensitive to amikacin, chloramphenicol, ciprofloxacin, gentamicin and ceftiofur and the highest frequency of resistance was observed to tetracycline, and ampicillin (52.8% and 37.1% respectively). The phylogenetic groups were identified by conventional triplex PCR. Results showed that E. coli strains segregated mainly in phylogenetic group B1, 52.8% in diarrheic herds and 52.9% in healthy herds.

Relationship between external and internal udder and teat measurements of machine milked dromedary camels.

This study aims to determine the relationship between internal and external udder and teat measurements and evaluate their correlation with milk yield and milk partitioning in the udder of dromedary camels. Six Maghrebi camels reared under intensive conditions were monitored at early, middle, and late lactation. Udder measurements included udder depth, udder horizontal circumference, fore and rear teat length and diameter. Besides, scans of the left fore and rear quarters were taken in duplicate before morning milking (16 h) using an oxytocin receptor blocking agent (Atosiban) to determine teat and gland distension before milk ejection. Cisternal and alveolar milk volumes were then evaluated. Correlation coefficients were calculated between the performed udder external and ultrasonographical measurements and cisternal and daily milk production on the measurement day. Significant effect of lactation stage was observed in all measured traits. All internal and external measurements decreased significantly at late lactation as well as cisternal and total milk yield. The quarter cisternal area averaged 16.3 ± 2.2 cm(2) and decreased about three times at late lactation compared to early and middle lactation. All external and internal measurements were positively and highly correlated (P < 0.001). The knowledge of the relationship between udder internal and external morphological traits would permit to predict udder cisternal storage capacity and can ultimately be adopted to improve milk storage capacity of dromedary camels.

Passive transfer of maternal immunity in the dromedary (Camelus dromedarius), involvement of heavy chain antibodies.

In many mammalian species, newborns are agammaglobulinemic; thus, colostrum and milk are the main sources of early protective antibodies. These antibodies are produced in the mother's serum and transferred to mammalian glands a few days before parturition. Here, we have studied the transfer of immunity from a she-camel immunized with human serum albumin (HSA) to her calf via colostrum and milk. Our results show that HSA-specific antibodies are produced in the mother's serum and are subsequently transferred to her colostrum. These specific antibodies are then transferred by suckling to the calf. The calf serum did not contain HSA-reactive antibodies at parturition and before the first feed, after suckling, a rise in reactivity was observed peaking at 24 h postpartum. The involvement of heavy chain antibodies (HCAbs) in the process of immunity transfer was also examined, and it was found that they were also transferred from the colostrum to the calf serum like conventional antibodies.